LOCUS       V018987                 5910 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018987
VERSION     V018987
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        1..1179
                     /note="strong constitutive promoter for human elongation
                     factor EF-1α"
                     /label="EF-1α promoter"
     promoter        1196..1214
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     protein_bind    1293..1317
                     /bound_moiety="BP Clonase™"
                     /gene="mutant version of&nbsp;attB"
                     /note="recombination site for the Gateway® BP reaction"
                     /label="attB1"
     CDS             1322..1780
                     /label="EIF5A2(NM_020390)"
                     /note="EIF5A2(NM_020390)"
                     /gene="EIF5A2"
     CDS             1781..1804
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     polyA_signal    1838..2062
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     protein_bind    2132..2165
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
                     /label="loxP"
     primer_bind     complement(2623..2639)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     promoter        3113..3217
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             3218..4078
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     rep_origin      4249..4837
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    5125..5146
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        5161..5191
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5199..5215
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     primer_bind     5223..5239
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5737..5770
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
                     /label="loxP"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtactaata
     1201 cgactcacta tagggagacc caagctggct aggtaagctt ggtaccgagc tcggatccac
     1261 tagtccagtg tggtggaatt ctgcagatat caacaagttt gtacaaaaaa gcaggctttc
     1321 aatggcagac gaaattgatt tcactactgg agatgccggg gcttccagca cttaccctat
     1381 gcagtgctcg gccttgcgca aaaacggctt cgtggtgctc aaaggacgac catgcaaaat
     1441 agtggagatg tcaacttcca aaactggaaa gcatggtcat gccaaggttc accttgttgg
     1501 aattgatatt ttcacgggca aaaaatatga agatatttgt ccttctactc acaacatgga
     1561 tgttccaaat attaagagaa atgattatca actgatatgc attcaagatg gttacctttc
     1621 cctgctgaca gaaactggtg aagttcgtga ggatcttaaa ctgccagaag gtgaactagg
     1681 caaagaaata gagggaaaat acaatgcagg tgaagatgta caggtgtctg tcatgtgtgc
     1741 aatgagtgaa gaatatgctg tagccataaa accctgcaaa gattacaagg atgacgacga
     1801 taagtgacca tggtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     1861 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     1921 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     1981 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     2041 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     2101 tatccccacg cgccctgtag cggcgcatta aataacttcg tataatgtat gctatacgaa
     2161 gttatatcgt atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
     2221 accctaactg acacacattc cacagaattc cttgaaatac atccacaggt acacctccaa
     2281 ttgactcaaa tgatgtcaat tagtctatca gaagcttcta aagccatgac atcattttct
     2341 ggaattttcc aagctgttta aaggcacagt caacttagtg tatgtaaact tctgacccac
     2401 tggaattgtg atacagtgaa ttataagtga aataatctgt ctgtaaacaa ttgttggaaa
     2461 aatgacttgt gtcatgcaca aagtagatgt cctaactgac ttgccaaaac tattgtttgt
     2521 taacaagaaa tttgtggagt agttgaaaaa cgagttttaa tgactccaac ttaagtgtat
     2581 gtaaacttcc gacttcaact gtagggtacc gagctcgaat tcactggccg tcgttttaca
     2641 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     2701 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     2761 cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     2821 ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     2881 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     2941 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     3001 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     3061 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     3121 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3181 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     3241 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     3301 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     3361 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     3421 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     3481 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     3541 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     3601 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     3661 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     3721 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     3781 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     3841 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     3901 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     3961 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     4021 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     4081 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4141 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4201 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4261 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4321 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4381 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4441 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4501 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4561 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4621 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4681 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4741 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4801 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4861 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     4921 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     4981 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     5041 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     5101 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     5161 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     5221 cacaggaaac agctatgacc atgattacgc caagcttgca tgcctgcagg tcgactctag
     5281 acctatacag ttgaagtcgg aagtttacat acacttaagt tggagtcatt aaaactcgtt
     5341 tttcaactac tccacaaatt tcttgttaac aaacaatagt tttggcaagt cagttaggac
     5401 atctactttg tgcatgacac aagtcatttt tccaacaatt gtttacagac agattatttc
     5461 acttataatt cactgtatca caattccagt gggtcagaag tttacataca ctaagttgac
     5521 tgtgccttta aacagcttgg aaaattccag aaaatgatgt catggcttta gaagcttctg
     5581 atagactaat tgacatcatt tgagtcaatt ggaggtgtac ctgtggatgt atttcaaggg
     5641 gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagcgg
     5701 ccgctagctc gagatcttaa ttaaactagt atcgatataa cttcgtataa tgtatgctat
     5761 acgaagttat atgtacgggc cagatatacg cgttgacatt gattattgac taggcttttg
     5821 caaaaagctt tgcaaagatg gataaagttt taaacagaga ggaatctttg cagctaatgg
     5881 accttctagg tcttgaaagg agtgggaatt
//
